SNT 1200-2003 烟草中转基因成分定性PCR检测方法

ProtocolofqualitativePCRforthedetectionofgeneticallymodifiedcomponentsintobaccoa3=.3.2.13TrisItris(hydroxymethyl)arninomethane,Z(%4S)JfilSEflti2a3.2.14DNAIde~x~ribonucleicacid,li##@#$I@.3.2.15dNTPs1dTTP,dATP,dCTP,dGTP.3.2.16dTTPIdeoxyribonucleosidetriphosphate,M#@VZ#l@3.2.17dATPldeoxyadenosinetriphosphate,ll#@BP3@Ra3.2.18dCTPldeoxycytidlnetriphosphate,W#lekB3fEl#l@.3.2.19dGTPldeoxyguanosinekphosphate,%%4~~@@3.2.20dUTPIdeoxyuridinetriphoaphate,lR#R%Z119B!.3.2.21bptbasepair,&&Xlf.fQaH&#s&At7SH##&&#tlP#$d@&W7&sb.k9'rcH&dBflBA,#&BJ&$$8.$!N@###aH&&*&?@&&@#BekQ,#&.&@&#,BR*&&fl,P;f*I%#.RH&3244fi~&~Rs~&a~~&&~~~*Am~*&~~~~mHm~m~m~~&~~~s&~~#~~~sw~~R-+%sm~B~~~~~#+~~~~B~~~~k?uA~#~m~~~&%%~fiRMB#&ESHaHRSB@BM&N&SHB@HH@@nBaF*&k?,B#BeSMSfi&B~##ESMbC#LiSH&;J$BBUWaN'F#SBM5619S8'H83iM@J3=91%i35sG&?,NOs@kPNB#SESB;NPT-I1r'#!&!,i$lbbkrtfal$&IEI&SiATMV-CP,CMV-CP,TMV-54KD,RT,PVY-CP.6.1*B?@miH$&M8GB/T668243~~7JC(Z%*)M)5#5.6.2-Fn~Bl~*~jifQ~#~~~~~~~N~6,2.1akB,Z~~B.#r#gt,CTAB,%%,EDTA,Tris,BQ@,P%~iFiLe6.2.2dNTPa*TaqDNAfltf3sBC,bl~#f~t#~ZBi!,DNAj).?.it6lE.&%4&#@?4%.6.3X#ZPE@Ylllkn(%%1).1BR90SBatarllA,*ZRS~IJiR3I#Afla~126R35sNOSNPT-I1BtCMVCP91¶b@PJ5'-gctcctacaaatgccatca-3'5'-grtagtgggattgtgcgtca-3'5'-gaatcctgttgccggtcttg3'.5'-ttatcctagtttgcgcgcta-3'5'-ggatctcctgtcatct-3'5'-gatcatcctgatcgac-3'5'-cc~actagttaacaatttgatt~ga-3'-5'-ccggaag~tttaggact~ta~gt-3'5'-aagacgttggcagctggtcg3'5'-ctcgaatttgaatgcgcgaa-3'&851?RH9Cn)hi#9bMfimmuFtll#&.tf;B19%~l8obp173bp299bp202bpbB%Blk39t;llW%B!hlMIY1111%Yr$l!&MPS!SbtlTrisItris(hydroxymethyl)aminomethane,3(%qgoDNAIdeoxyribonucleicacid,li##.$I###@.dNTPsIdTTP,dATP,dCTP,dGTP.dTTPIdeoxyribonucleoaidetriphosphate,NRll$%f:3@@.dATPIdeoxyadenosinetriphosphate,j3jt%#l$[e-mail]sphate,W#$Mft:[e-mail]*iphosphate,W#&$P3@@.dUTPsdeoxyuridinetriphosphate,N#R~G~@.bp,basepair,MSX$.R~E~Q%~~T$BrH#WBrJ~&~PR*~~SH&+~BE1SBB~3A~XSHS+.$QbENSiri&J)*#SBBQ*%?@#C@1I#C#i,%&\%bW%l\RR#C&iB,Pf%tk#*EU&%+?a@~R~B~a~~~~~~mBAmU*B~~~~mHM~M~laJMs&~m$~~~~fi~BMSH~8%-+%SHS~M&%R*#42~&~B3F~%k~U2%fi~~~2~B%RHfi~M3&##~S~*~Rsm&BH@w&~a~*Nm**9JB3F,%kF,%#~z&a3fiaB*#BRNR#MHMSBUfoRI'feQSBn~%#HMMBJYPjjm35SG3F,NOS%JkFjjll1$#6ESBNPT-I1r'#!4&i@BfiBlfEI&&BB$jTMV-CP,CMV-CP,TMV-54KD.RT,PVY-CP.6.1~B?BmiH*&#&GB/T6682EP~%dC(3#*)#3!4#.6.2-FNlje#PlB*gift#e%fi**%?*fi%HtpJ*6.2.1ak#.3~)&Efl,%&tft,CTAB,%Pi.811,EDTA.Tris,lJQ@.~%&FiZBe.6.2.2dNTPs,TaqDNAflt&BBC,~l~M~~fiZ@.DNA%F.it68€.$3dr~#$il#%.6.3XH3BBB3l4@(%$!S1).11W9PiiBetsrVA.9E@iBIJiR3I#AN*&tda~Bl35sNOSNPT-11BtCMVCP91488;945'-gctcctacaaatgccatca-3'5'-gatagtgggattgtgcgtca-315'-gaatcctgttgccggtcttg3'5'-ttatcctagtttgcgcgcta-3'5'-ggatctcctgtcatct-3'5'-gatcntcctgatcgac-3'5'-cccactagttaacaatttgnttgga-3'5'-ccg~aagctttaggactgtaggt-3'5'-aagacgttggcagctggtcg-3'5'-ctcgbatttgaat~cgcgu-3'*88?4€M9t.a9t.m9bt!lfim4'tdMtrN#&Eb195bpleobp173bp299bp202hp83%Bm&#z815RBW!!l%&Y1RPgBMP&UI7.1%%@.7.2PCRN.7.3@M%QiYR.7.4#la&$IIRlG&.75*%+A.76l$tlPll&#%%fE.77&*I#i!?a7.8Q88f.79&E%'Bb14W1.7.10WPT#BL.711%7k#.712&fi##$$j@##(O.5pL.-IOOOpI-),+'$A#fts;(IOOPI.-1OOOILL),&3#~#$~L~~(250pL-1500pl.),@f#$.8.1W#BbilWSN/T1194qB%H-fi&8?io8.2(8]##fiURB%~;t$l~t*;t$lN&~%l~EXi8:AtljFb([8:(~~HR~i.13~;~JR~~%!%IBIJ@&BIJ%~IIEfi,L27C-28CtPC%#)&ji~~-k*S,a;trIrH#NKl#NRA#al~~t-~ZoomgYfT@&EXB?8src:~W#h*Ltt;#@B9YfiM=EZ~*HBfiS~~@*tRfib~A~4t%&*%BjrrfiWB~@at-#&~;W.8.3DNAH#!&83.1&@@?#I%DNAB(J~~*%iSt2~Cl~ifld*4%flilSBcJ~#tk~&BfiDNA@@.8.32#FfjFll&#@lBt+DNA&I@%.8.3.21#~i)i~%~~~t&@HliBll%@WFEGAC1%)ik%%91l15m1.Rlh4F'P,#$IS%1pI./rngbUA65Cm&&l~xXI'AB#~~~~P~P%~~Z~&!AM~A)*%Z%(#W3)B)fiIIAP-%SZz,BP,R3)R*J,65C;r)C%irtt-bn$&1tn~n--3min,Ltt;?&+tkL!lIftM,Mf!!#l~t-NB~.8.3.22h~A$,#&IXI3.~Wfr/#fift(24l,#8!3)&),$@&%,@&a&,13OOOr/m~n,L85min.31l12&81PVY-CPTMV54KDTMV-CP---Chloroplast~~on-coadingDNAtrtl)#BQEB362bp295ph327b~550bp15%$!&till#Z#iMgnushI#Rl(SB4€1IR+I-m,$1infimha$1I?Jd51-gatalttcaestaclcgeeca-3'5'-gcataacgcg~taaacc~ne3'5'-gagttgtct~1(c~tcattga-3'5'-acaatggtcaasgccgggta-3'--5'-~tg~tctt~tca~cagcg~gggc-3'5'-caccgttgcgtcgtcttrcrctacg-3'----.5'-cgeaatcggtagacgcracg-3'5'-ggggaregegggactrpaae-3'8.3.2.3#LE*~~A-#&RJL~%~v~~@ATR%#~BuA~~)X-#~~10%CTAB(65tWB),a$=&fRA3#RHG%FB/%aM(2411,#@3)~323)?l3~,13OOOr/minFI(IJ~5min.lO%CTABa)r/lfisCJ&#!l%NI#%kA.8.3.2.4#k_f:*a%saia--%JistJ~LviP'~ruAqiiHHCTAB~s~mt,ssm~,15000r/minff~L10min,%E#B.CTABE$i!&!)r/l%l&!#J#!&gijkA.8.3.2.5$RE+fRA200rL%&kTE@#~,65CB#310minp###25i@.##!Em#A.8.3.2.6bUA=@.gb;lFRb)(F?MtBpV;5000r/rninat10rnin,Srl.Ej$$$m.8.3.2.7M8o%Zs5r%%B~-M~3g9~T&hS%T~PR6A~fi3)~%&1.t.M~~@H~B~DNA%BjeoDzw/0480ttI;.Mt?Z11.7-1.9kM@fl,RePCRI&NS!r)t.8.3.3T#lw-B#9a*DNAM&%8.3.3.1#8.2+i@lflM@#5iC%100me%h-%h!J1.5m1.a~b'l!F+,fin700pI.1XCTARM&@Fb313itEi#l%%,65cPB10min.0.3.3.2fiAA~#$R5$IYB/FF&lS(24l,#R~B$$3%l!&Hj#r913OOOr/minstJ~t5min.8.3.3.3IQ~~R&;~SI%A-%~~KIRJL$T~P'*fi~AffiX-#%JIlo%CTABRtlBB€#%,sE/Q,SbnA7B;#$R=gl@%-WBB(241,&B~)B,%3)?~&,13000r/minRJ~5min.8.3.3.4%kElk~41+-%iliH#~L'14:+~BZI:-$~#Sb;li~A~@HCTABtKB#%li#i,15000r/min,R~t10rnin,B@@@.8.3.3.5&Ei@iP'fitlA200pLB&kTEHFb@,65CB%10rnin,#@@%Re8.3.3.6bnA=f$f@HBkF?HZiI$,-70c#4$30minV15000r/min10min,~~@#~.